Poly(dimethylsiloxane) (PDMS) capillary electrophoresis (CE) microchips were modified by a dynamic coating method that provided stable electroosmotic flow (EOF) with respect to pH. The separation channel was coated with a polymer bilayer consisting of a cationic layer of Polybrene (PB) and an anionic layer of dextran sulfate (DS). According to the difference in charge, PB- and PB/ DS-coated channels supported EOF in different directions; however, both methods of channel coating exhibited a pH-independent EOF in the pH range of 5-10 due to chemical control of the effective zeta-potential. The endurance of the PB-coated layer was determined to be 50 runs at pH 3.0, while PB/DS-coated chips had a stable EOF for more than 100 runs. The effect of substrate composition and chip-sealing methodology was also evaluated. All tested chips showed the same EOF on the PB/DS-coated channels, as compared to uncoated chips, which varied significantly. No significant variation for separation and electrochemical detection of dopamine and hydroquinone between coated and uncoated channels was observed.